title: Immobilized butyrylcholinesterase in the characterization of new inhibitors that could ease Alzheimer's disease.
abstract: Focus of this work was the development and characterization of a new immobilized enzyme reactor (IMER) containing human recombinant butyrylcholinesterase (rBChE) for the on-line kinetic characterization of specific, pseudo-irreversible and brain-targeted BChE inhibitors as potential drug candidates for Alzheimer's disease (AD). Specifically, a rBChE-IMER containing 0.99 U of covalently bound target enzyme was purposely developed and inserted into a HPLC system connected to a UV-vis detector. Selected reversible cholinesterase inhibitors, (-)-phenserine and (-)-cymserine analogues, were then kinetically characterized by rBChE-IMER, and by classical in solution assays and their carbamoylation and decarbamoylation constants were determined. The results support the elucidation of the potency, inhibition duration, mode of action and specific structure/activity relations of these agents and allow cross-validation of the two assay techniques.
authors.affiliation: Department of Pharmaceutical Sciences, Via Belmeloro 6, University of Bologna, 40126 Bologna, Italy.
authors.forename: Manuela
authors.initials: M
authors.lastname: Bartolini
authors.forename: Nigel H
authors.initials: NH
authors.lastname: Greig
authors.forename: Qian-Sheng
authors.initials: QS
authors.lastname: Yu
authors.forename: Vincenza
authors.initials: V
authors.lastname: Andrisano
mesh_terms: D000544:Alzheimer Disease / Q000188:drug therapy*; D002091:Butyrylcholinesterase / Q000737:chemistry* / Q000187:drug effects; D002800:Cholinesterase Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D004353:Drug Evaluation, Preclinical; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000187:drug effects; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry / Q000187:drug effects
chemical_list: D002800:Cholinesterase Inhibitors; D004800:Enzymes, Immobilized; D011994:Recombinant Proteins; D002091:Butyrylcholinesterase